Genetic reprogramming with stem cells regenerates glomerular epithelial podocytes in Alport syndrome

Valerie S. Lebleu; Keizo Kanasaki; Sara Lovisa; Joseph L. Alge; Jiha Kim; Yang Chen; Yingqi Teng; Behzad Gerami-Naini; Hikaru Sugimoto; Noritoshi Kato; Ignacio Revuelta; Nicole Grau; Jonathan P. Sleeman; Gangadhar Taduri; Akane Kizu; Shahin Rafii; Konrad Hochedlinger; Susan E. Quaggin; Raghu Kalluri

doi:10.26508/lsa.202402664

Genetic reprogramming with stem cells regenerates glomerular epithelial podocytes in Alport syndrome

Valerie S. Lebleu^*, Keizo Kanasaki, Sara Lovisa, Joseph L. Alge, Jiha Kim, Yang Chen, Yingqi Teng, Behzad Gerami-Naini, Hikaru Sugimoto, Noritoshi Kato, Ignacio Revuelta, Nicole Grau, Jonathan P. Sleeman, Gangadhar Taduri, Akane Kizu, Shahin Rafii, Konrad Hochedlinger, Susan E. Quaggin, Raghu Kalluri^*

^*Corresponding author for this work

Medicine, Nephrology Division

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

Abstract

Glomerular filtration relies on the type IV collagen (ColIV) network of the glomerular basement membrane, namely, in the triple helical molecules containing the α3, α4, and α5 chains of ColIV. Loss of function mutations in the genes encoding these chains (Col4a3, Col4a4, and Col4a5) is associated with the loss of renal function observed in Alport syndrome (AS). Precise understanding of the cellular basis for the patho-mechanism remains unknown and a specific therapy for this disease does not currently exist. Here, we generated a novel allele for the conditional deletion of Col4a3 in different glomerular cell types in mice. We found that podocytes specifically generate α3 chains in the developing glomerular basement membrane, and that its absence is sufficient to impair glomerular filtration as seen in AS. Next, we show that horizontal gene transfer, enhanced by TGFβ1 and using allogenic bone marrow–derived mesenchymal stem cells and induced pluripotent stem cells, rescues Col4a3 expression and revive kidney function in Col4a3-deficient AS mice. Our proof-of-concept study supports that horizontal gene transfer such as cell fusion enables cell-based therapy in Alport syndrome.

Original language	English (US)
Article number	e202402664
Journal	Life science alliance
Volume	7
Issue number	6
DOIs	https://doi.org/10.26508/lsa.202402664
State	Published - Jun 2024

Funding

This work was supported by NIH Grant DK55001. The Kalluri laboratory was also supported by research funds from the MDACC and Beth Israel Deaconess Medical Center for the Division of Matrix Biology. VS LeBleu received funding from the NIH Research Training Grant in Gastroenterology (2T32DK007760-11) and the National Kidney Foundation of Illinois Medical Student Research Grant Award to Feinberg School of Medicine, Northwestern University, Chicago, IL. H Sugimoto received funding from the NIH Research Training Grant in Cardiovascular Medicine (5T32HL007374-30), and I Revuelta is funded by the Spanish Society of Nephrology. JP Sleeman received funding from the Deutsch Forschungsgemeinschaft (DFG) under the auspices of the D-A-CH project STOP-CKD, project number 492766284 (SL 37 12-1). We wish to thank the laboratory of Dr. Martin Pollak (Beth Israel Deaconess Medical Center, Boston) for teaching us how to perform bead extraction of glomeruli, Dr. Caroline A. Miller (Indiana University School of Medicine, Indianapolis) for her assistance with electron microscopy data capture, and Dr. Rolf Bjerkvig (University of Bergen, Norway) for his helpful discussions regarding cell fusion-related experiments. We wish to thank Dr. Joyce O\u2019Connell and Lizzy Finan for technical assistance and help in generating the figures and Kenneth Dunner and the High-Resolution Electron Microscopy Facility supported by CCSG grant NIH P30CA016672. This work was supported by NIH Grant DK55001. The Kalluri laboratory was also supported by research funds from the MDACC and Beth Israel Deaconess Medical Center for the Division of Matrix Biology. VS LeBleu received funding from the NIH Research Training Grant in Gastroenterology (2T32DK007760-11) and the National Kidney Foundation of Illinois Medical Student Research Grant Award to Feinberg School of Medicine, North-western University, Chicago, IL. H Sugimoto received funding from the NIH Research Training Grant in Cardiovascular Medicine (5T32HL007374-30), and I Revuelta is funded by the Spanish Society of Nephrology. JP Sleeman received funding from the Deutsch Forschungsgemeinschaft (DFG) under the auspices of the D-A-CH project STOP-CKD, project number 492766284 (SL 37 12-1). We wish to thank the laboratory of Dr. Martin Pollak (Beth Israel Deaconess Medical Center, Boston) for teaching us how to perform bead extraction of glomeruli, Dr. Caroline A. Miller (Indiana University School of Medicine, Indianapolis) for her assistance with electron microscopy data capture, and Dr. Rolf Bjerkvig (University of Bergen, Norway) for his helpful discussions regarding cell fusion-related experiments. We wish to thank Dr. Joyce O\u2019Connell and Lizzy Finan for technical assistance and help in generating the figures and Kenneth Dunner and the High-Resolution Electron Microscopy Facility supported by CCSG grant NIH P30CA016672.

ASJC Scopus subject areas

Ecology
Biochemistry, Genetics and Molecular Biology (miscellaneous)
Plant Science
Health, Toxicology and Mutagenesis

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10.26508/lsa.202402664

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Lebleu, V. S., Kanasaki, K., Lovisa, S., Alge, J. L., Kim, J., Chen, Y., Teng, Y., Gerami-Naini, B., Sugimoto, H., Kato, N., Revuelta, I., Grau, N., Sleeman, J. P., Taduri, G., Kizu, A., Rafii, S., Hochedlinger, K., Quaggin, S. E., & Kalluri, R. (2024). Genetic reprogramming with stem cells regenerates glomerular epithelial podocytes in Alport syndrome. Life science alliance, 7(6), Article e202402664. https://doi.org/10.26508/lsa.202402664

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title = "Genetic reprogramming with stem cells regenerates glomerular epithelial podocytes in Alport syndrome",

abstract = "Glomerular filtration relies on the type IV collagen (ColIV) network of the glomerular basement membrane, namely, in the triple helical molecules containing the α3, α4, and α5 chains of ColIV. Loss of function mutations in the genes encoding these chains (Col4a3, Col4a4, and Col4a5) is associated with the loss of renal function observed in Alport syndrome (AS). Precise understanding of the cellular basis for the patho-mechanism remains unknown and a specific therapy for this disease does not currently exist. Here, we generated a novel allele for the conditional deletion of Col4a3 in different glomerular cell types in mice. We found that podocytes specifically generate α3 chains in the developing glomerular basement membrane, and that its absence is sufficient to impair glomerular filtration as seen in AS. Next, we show that horizontal gene transfer, enhanced by TGFβ1 and using allogenic bone marrow–derived mesenchymal stem cells and induced pluripotent stem cells, rescues Col4a3 expression and revive kidney function in Col4a3-deficient AS mice. Our proof-of-concept study supports that horizontal gene transfer such as cell fusion enables cell-based therapy in Alport syndrome.",

author = "Lebleu, {Valerie S.} and Keizo Kanasaki and Sara Lovisa and Alge, {Joseph L.} and Jiha Kim and Yang Chen and Yingqi Teng and Behzad Gerami-Naini and Hikaru Sugimoto and Noritoshi Kato and Ignacio Revuelta and Nicole Grau and Sleeman, {Jonathan P.} and Gangadhar Taduri and Akane Kizu and Shahin Rafii and Konrad Hochedlinger and Quaggin, {Susan E.} and Raghu Kalluri",

note = "Publisher Copyright: {\textcopyright} 2024 LeBleu et al.",

year = "2024",

month = jun,

doi = "10.26508/lsa.202402664",

language = "English (US)",

volume = "7",

journal = "Life science alliance",

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Lebleu, VS, Kanasaki, K, Lovisa, S, Alge, JL, Kim, J, Chen, Y, Teng, Y, Gerami-Naini, B, Sugimoto, H, Kato, N, Revuelta, I, Grau, N, Sleeman, JP, Taduri, G, Kizu, A, Rafii, S, Hochedlinger, K, Quaggin, SE & Kalluri, R 2024, 'Genetic reprogramming with stem cells regenerates glomerular epithelial podocytes in Alport syndrome', Life science alliance, vol. 7, no. 6, e202402664. https://doi.org/10.26508/lsa.202402664

TY - JOUR

T1 - Genetic reprogramming with stem cells regenerates glomerular epithelial podocytes in Alport syndrome

AU - Lebleu, Valerie S.

AU - Kanasaki, Keizo

AU - Lovisa, Sara

AU - Alge, Joseph L.

AU - Kim, Jiha

AU - Chen, Yang

AU - Teng, Yingqi

AU - Gerami-Naini, Behzad

AU - Sugimoto, Hikaru

AU - Kato, Noritoshi

AU - Revuelta, Ignacio

AU - Grau, Nicole

AU - Sleeman, Jonathan P.

AU - Taduri, Gangadhar

AU - Kizu, Akane

AU - Rafii, Shahin

AU - Hochedlinger, Konrad

AU - Quaggin, Susan E.

AU - Kalluri, Raghu

PY - 2024/6

Y1 - 2024/6

N2 - Glomerular filtration relies on the type IV collagen (ColIV) network of the glomerular basement membrane, namely, in the triple helical molecules containing the α3, α4, and α5 chains of ColIV. Loss of function mutations in the genes encoding these chains (Col4a3, Col4a4, and Col4a5) is associated with the loss of renal function observed in Alport syndrome (AS). Precise understanding of the cellular basis for the patho-mechanism remains unknown and a specific therapy for this disease does not currently exist. Here, we generated a novel allele for the conditional deletion of Col4a3 in different glomerular cell types in mice. We found that podocytes specifically generate α3 chains in the developing glomerular basement membrane, and that its absence is sufficient to impair glomerular filtration as seen in AS. Next, we show that horizontal gene transfer, enhanced by TGFβ1 and using allogenic bone marrow–derived mesenchymal stem cells and induced pluripotent stem cells, rescues Col4a3 expression and revive kidney function in Col4a3-deficient AS mice. Our proof-of-concept study supports that horizontal gene transfer such as cell fusion enables cell-based therapy in Alport syndrome.

AB - Glomerular filtration relies on the type IV collagen (ColIV) network of the glomerular basement membrane, namely, in the triple helical molecules containing the α3, α4, and α5 chains of ColIV. Loss of function mutations in the genes encoding these chains (Col4a3, Col4a4, and Col4a5) is associated with the loss of renal function observed in Alport syndrome (AS). Precise understanding of the cellular basis for the patho-mechanism remains unknown and a specific therapy for this disease does not currently exist. Here, we generated a novel allele for the conditional deletion of Col4a3 in different glomerular cell types in mice. We found that podocytes specifically generate α3 chains in the developing glomerular basement membrane, and that its absence is sufficient to impair glomerular filtration as seen in AS. Next, we show that horizontal gene transfer, enhanced by TGFβ1 and using allogenic bone marrow–derived mesenchymal stem cells and induced pluripotent stem cells, rescues Col4a3 expression and revive kidney function in Col4a3-deficient AS mice. Our proof-of-concept study supports that horizontal gene transfer such as cell fusion enables cell-based therapy in Alport syndrome.

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DO - 10.26508/lsa.202402664

M3 - Article

C2 - 38561223

AN - SCOPUS:85189827643

SN - 2575-1077

VL - 7

JO - Life science alliance

JF - Life science alliance

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M1 - e202402664

ER -

Genetic reprogramming with stem cells regenerates glomerular epithelial podocytes in Alport syndrome

Abstract

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